We have cloned an actin-encoding cDNA from the dimorphic fungus Histoplasma capsulatum, an important pathogen of humans. The predicted amino acid sequence as well as the general codon pattern of Histoplasma actin revealed the highest degree of similarity to the actin of the filamentous ascomycete Aspergillus nidulans. Southern blot analysis determined that actin was encoded by a single copy in the Histoplasma genome. Northern blot analysis showed a single 1700 nt transcript in yeast and mould cells as well as in cells undergoing the temperature induced mould-to-yeast conversion. Actin mRNA levels normalized to 18 S rRNA were found to be equivalent in all the stages examined, except for a sharp four-fold transient decrease 4 h into the mould-to-yeast conversion. These data suggest that actin mRNA would not be a suitable internal marker for expression studies during Histoplasma mould-to-yeast morphogenesis.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1080/02681219780001091 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Twelve actin-encoding cDNAs from the American lobster, Homarus americanus: cloning and tissue expression of eight skeletal muscle, one heart, and three cytoplasmic isoforms.
Comp Biochem Physiol B Biochem Mol Biol
June 2009
Department of Marine Biology, Pukyong National University, Busan, 608-737 South Korea.
Lobster muscles express a diverse array of myofibrillar protein isoforms. Three fiber types (fast, slow-twitch or S1, and slow-tonic or S2) differ qualitatively and quantitatively in myosin heavy and light chains, troponin-T, -I, and -C, paramyosin, and tropomyosin variants. However, little is known about the diversity of actin isoforms present in crustacean tissues.
View Article and Find Full Text PDFIsolation and characterization of a single-copy actin gene from a sterile mutant of Ulva pertusa (Ulvales, Chlorophyta).
Gene
June 2004
Laboratory of Marine Biochemistry, Faculty of Bioresources, Mie University, Tsu Mie 514-8507, Japan.
We constructed a cDNA library from sterile Ulva pertusa (Ulvales, Chlorophyta), and isolated and characterized a full-length cDNA clone encoding actin. The actin (ACT) cDNA consisted of 1487 nucleotides (nt) and had an open reading frame (ORF) encoding a polypeptide of 377 amino acid (AA) residues. The ACT gene had one intron in the 5'-untranslated region and three introns in the coding region.
View Article and Find Full Text PDFIsolation of actin-encoding cDNAs from symbiotic corals.
DNA Res
December 2002
Division of Marine Biosciences, Ocean Research Institute, The University of Tokyo, 1-15-1 Minamidai, Nakano, Tokyo 164-8639, Japan.
A cDNA (named LGfact) encoding actin was identified in planular larvae of the scleractinian coral Galaxea fascicularis, using the reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) techniques. RNA from the adult coral that was inhabited by symbiotic dinophytes was subjected to a similar RT-PCR, and a cDNA fragment, named AGfact-p, was found to encode an actin form distinct from LGfact. In an expression study, LGfact transcripts were present at similar levels in asymbiotic larvae and symbiotic adults, indicating that LGfact was expressed by the host.
View Article and Find Full Text PDFMolecular characterization of actin genes from homobasidiomycetes: two different actin genes from Schizophyllum commune and Suillus bovinus.
Gene
June 2000
Department of Biosciences, Division of Plant Physiology, University of Helsinki, Finland.
The actin-encoding genes Scact1 and Scact2 of the homobasidiomycete Schizophyllum commune are the first actin genes isolated from higher filamentous fungi. Their isolation shows that homobasidiomycetes have two actin encoding genes instead of one typical to yeasts and filamentous ascomycetes. This result was further confirmed by cloning two actin encoding genes, Sbact1 and Sbact2, from another homobasidiomycete Suillus bovinus.
View Article and Find Full Text PDFThe stress kit: a new method based on competitive reverse transcriptase-polymerase chain reaction to quantify the expression of human alphaB-crystallin, Hsp27, and Hsp60.
Cell Stress Chaperones
January 2000
Division of Immunological and Infectious Diseases, TNO Prevention and Health, Leiden, The Netherlands.
We describe a reverse transcriptase-polymerase chain reaction method for the semiquantitative detection of mRNAs encoding the human heat shock proteins alphaB-crystallin, Hsp27, and Hsp60. The method involves the coamplification of cellular mRNA-derived cDNA with a dilution series of a competitor fragment (internal standard), using 1 primer pair common to both templates. Internal standards were based on cellular-derived cDNA engineered to be slightly smaller to differentiate between the target and the standard on electrophoretic separation.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!